Basement Membrane of Small Diameter Xenogeneic Extracellular Matrix Scaffolds Modulate Quiescent Human Endothelial Cell Monolayer Formation

2021 ◽  
Author(s):  
Manuela Lopera Higuita ◽  
Nicholas A. Shortreed ◽  
Surendra Dasari ◽  
Leigh Griffiths
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cell ◽  
Basement Membrane ◽  
Cell Monolayer ◽  
Small Diameter ◽  
Human Endothelial Cell ◽  
Endothelial Cell Monolayer ◽  
Monolayer Formation ◽  
Extracellular Matrix Scaffolds

scholarly journals Endothelial cell monolayer formation on a small-diameter vascular graft surface under pulsatile flow conditions

2021 ◽  
Vol 23 (3) ◽  
pp. 101-114
Author(s):  
M. Yu. Khanova ◽  
E. A. Velikanova ◽  
V. G. Matveeva ◽  
E. O. Krivkina ◽  
T. V. Glushkova ◽  
...  
Keyword(s):  
Shear Stress ◽  
Endothelial Cell ◽  
Pulsatile Flow ◽  
Vascular Graft ◽  
Cell Monolayer ◽  
Small Diameter ◽  
Flow Conditions ◽  
Endothelial Cell Culture ◽  
Monolayer Formation ◽  
Whole Transcriptome

Objective: to create a cell-populated small-diameter vascular graft (SDVG) using autologous endothelial cells and extracellular matrix proteins, and to evaluate the efficiency of endothelial cell monolayer formation during shear stress preconditioning in a SDVG.Materials and methods. PHBV/PCL tubular scaffolds of vascular grafts were made by electrospinning from a mixture of polyhydroxybutyrate-valerate (PHBV) copolymer and polycaprolactone (PCL) and modified with fibrin. To populate the graft, an endothelial cell culture was isolated from the blood of patients with coronary heart disease. Phenotyping of endothelial colony-forming cell (ECFC) culture was performed by flow cytometry and immunofluorescence microscopy. Cell proliferative and angiogenic activity were also studied. Cell-populated vascular scaffolds were cultured in a pulsatile flow setup with a final shear stress of 2.85 dyne/cm2. The effect of pulsatile flow on monolayer formation was assessed by immunofluorescence, scanning electron microscopy, atomic force microscopy, and whole-transcriptome RNA sequencing.Results. Under the influence of pulsatile flow, endothelial cells that were seeded into the tubular scaffold showed an increase in the expression level of endothelial profile proteins, focal adhesion and cytoskeleton. In contrast to endothelial cell culture on a vascular graft surface under static conditions, when cultured under pulsatile flow with 2.85 dyne/ cm2 shear stress, endothelial lining cells have an increased ability to adhere and are oriented along the pulsatile flow path. Whole-transcriptome RNA sequencing showed that induced shear stress increased expression levels of differentially expressed genes encoding proteins that ensure vascular development, endothelial integrity, and endothelial metabolism. A protocol for fabrication of a personalized cell-populated biodegradable SDVG under pulsatile flow conditions was developed.Conclusion. The use of autologous fibrin and ECFC culture, as well as shear stress preconditioning, allow to obtain a personalized cell-populated SDVG with continuous functional endothelial monolayer adapted to the flow.

Increase in permeability of human endothelial cell monolayer by recombinant human lymphotoxin

1989 ◽  
Vol 162 (3) ◽  
pp. 1431-1437 ◽  
Author(s):  
Katsuhiro Shinjo ◽  
Satoru Tsuda ◽  
Toshio Hayami ◽  
Takashi Asahi ◽  
Hajime Kawaharada
Keyword(s):  
Endothelial Cell ◽  
Cell Monolayer ◽  
Human Endothelial Cell ◽  
Endothelial Cell Monolayer

scholarly journals Bioinspired hydrogel surfaces to augment corneal endothelial cell monolayer formation

2021 ◽  
Author(s):  
Fatma Zehra Erkoc‐Biradli ◽  
Alp Ozgun ◽  
Meftune Özgen Öztürk‐Öncel ◽  
Merve Marcali ◽  
Caglar Elbuken ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Monolayer ◽  
Corneal Endothelial Cell ◽  
Endothelial Cell Monolayer ◽  
Monolayer Formation

scholarly journals Dystroglycan is selectively cleaved at the parenchymal basement membrane at sites of leukocyte extravasation in experimental autoimmune encephalomyelitis

2006 ◽  
Vol 203 (4) ◽  
pp. 1007-1019 ◽  
Author(s):  
Smriti Agrawal ◽  
Per Anderson ◽  
Madeleine Durbeej ◽  
Nico van Rooijen ◽  
Fredrik Ivars ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Basement Membrane ◽  
Experimental Autoimmune Encephalomyelitis ◽  
Cell Monolayer ◽  
Cerebral Vessels ◽  
Leukocyte Infiltration ◽  
Endothelial Cell Monolayer ◽  
Autoimmune Encephalomyelitis ◽  
Astrocyte Endfeet ◽  
Experimental Autoimmune

The endothelial cell monolayer of cerebral vessels and its basement membrane (BM) are ensheathed by the astrocyte endfeet, the leptomeningeal cells, and their associated parenchymal BM, all of which contribute to establishment of the blood–brain barrier (BBB). As a consequence of this unique structure, leukocyte penetration of cerebral vessels is a multistep event. In mouse experimental autoimmune encephalomyelitis (EAE), a widely used central nervous system inflammatory model, leukocytes first penetrate the endothelial cell monolayer and underlying BM using integrin β1-mediated processes, but mechanisms used to penetrate the second barrier defined by the parenchymal BM and glia limitans remain uninvestigated. We show here that macrophage-derived gelatinase (matrix metalloproteinase [MMP]-2 and MMP-9) activity is crucial for leukocyte penetration of the parenchymal BM. Dystroglycan, a transmembrane receptor that anchors astrocyte endfeet to the parenchymal BM via high affinity interactions with laminins 1 and 2, perlecan and agrin, is identified as a specific substrate of MMP-2 and MMP-9. Ablation of both MMP-2 and MMP-9 in double knockout mice confers resistance to EAE by inhibiting dystroglycan cleavage and preventing leukocyte infiltration. This is the first description of selective in situ proteolytic damage of a BBB-specific molecule at sites of leukocyte infiltration.

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